Xuebijing alters tumor necrosis factor-alpha, interleukin-1beta and p38 mitogen activated protein kinase content in a rat model of cardiac arrest following cardiopulmonary resuscitation

We established a rat model of cardiac arrest by clamping the endotracheal tube of adult rats at expiration. Twenty-four hours after cardiopulmonary resuscitation, nerve cell injury and expression of tumor necrosis factor-α, interleukin-1β, and p38 mitogen activated protein kinase content were increased. Rats injected with Xuebijing, a Chinese herb compound preparation, exhibited normal cellular structure and morphology, dense neuronal cytoplasm, and decreased tumor necrosis factor-α, interleukin-1β, and p38 mitogen activated protein kinase expression at 24 hours following cardiopulmonary resuscitation. These data suggest that Xuebijing can attenuate neuronal injury induced by hypoxia and reperfusion during cardiopulmonary resuscitation.

Altered pattern of tumor necrosis factor-alpha production in peripheral blood monocytes from Crohn's disease

AIM To evaluate the inflammatory state in Crohn's disease(CD) patients and correlate it with genetic background and microbial spreading.METHODS By means of flow cytometry, production of tumor necrosis factor-alpha(TNF-α) was measured in peripheral blood monocytes from patients suffering from CD, ulcerative colitis(UC) and in healthy subjects after stimulation of the NOD2 and TLR pathways. CD patients were genotyped for the three most common NOD2 variants(R702W, G908 R and L1007Pfs*2) and basal production of TNF-α was correlated to NOD2 genotype. Also, production of TNF-α was correlated to plasmatic levels of LPS Binding Protein(LBP), soluble(s) CD14 and to the activity state of the disease.RESULTS The patients with CD were characterized by a significantly higher monocyte basal expression of TNF-αcompared with healthy subjects and UC patients, and after stimulation with Pam3CSK4(ligand of TLR2/1) and MDP-L18(ligand of NOD2) this difference was maintained, while other microbial stimuli(LPS, ligand of TLR4 and Poly I:C, ligand of TLR3) induced massive activation in CD monocytes as well as in UC and in healthy control cells. There was no significant difference in the production of TNF- α between patients who carried CD-associated heterozygous or homozygous variants in NOD2 and patients with wild type NOD2 genotype. Although serum LBP levels have been shown to correlate positively with the state of activity of the disease, TNF-α production did not show a clear correlation with either LBP or s CD14 levels in plasma. Moreover, no clear correlation was seen between TNF-α production and activity indices in either CD or UC.CONCLUSION Peripheral monocytes from CD express higher basal and stimulated TNF-α than controls, regardless of NOD2 genotype and without a clear correlation with disease activity.

“Three Methods and Three Points” regulates p38 mitogen-activated protein kinase in the dorsal horn of the spinal cord in a rat model of sciatic nerve injury

Tuina is a traditional Chinese treatment for sensory disturbances caused by peripheral nerve injury and related diseases. Our previous studies showed that tuina regulates relevant regions and indices of the spinal dorsal horn using the Dian, Bo, and Rou method in Yinmen（BL37）, Yanglingquan（GB34）, and Weizhong（BL40）. Treatment prevents muscle atrophy, protects spinal cord neurons, and promotes sciatic nerve repair. The mechanisms of action of tuina for treating peripheral nerve injury remain poorly understood. This study established rat models of sciatic nerve injury using the crushing method. Rats received Chinese tuina in accordance with the principle of ＂Three Methods and Three Points,＂ once daily for 20 days. Tuina intervention reduced paw withdrawal latency and improved wet weight of the gastrocnemius muscle, as well as promoting morphological recovery of sciatic nerve fibers, Schwann cells, and axons. The protein expression levels of phospho-p38 mitogen-activated protein kinase, tumor necrosis factor-α, and interleukin-1β also decreased. These findings indicate that ＂Three Methods and Three Points＂ promoted morphological recovery and improved behavior of rats with peripheral nerve injury.

Perilla oil and exercise decrease expressions of tumor necrosis factor-α,plasminogen activator inhibitor-1 and highly sensitive C-reactive protein in patients with hyperlipidemia

OBJECTIVE:To verify the effects of perilla oil on the regulation of blood lipid levels in patients with hyperlipidemia.METHODS:Blood was taken from patients prior to and 8 weeks following treatment with perilla oil.Different ways to test for indexes which correlate to hyperlipidemia were performed.Some indexes,which correlate with inflammation and injury to endothelial cells,were tested using enzyme linked immunosorbent assays.RESULTS:Serum lipid levels [triglyceride(TG),total cholesterol(TC),and low-density lipoprotein-cholesterol(LDL-C)] changed significantly after 56 days of treatment.Differences were noted as early as 28 days after treatment began(P<0.05).Treatment with perilla oil showed statistically significant recovery levels of high-density lipoprotein-cholesterol(HDL-C) after 28 and 56 days of treatment.Plasma lipids levels were significantly lower after 56 days of treatment(P<0.05).Perilla oil reduced blood lipid levels in patients,and the regulation of cell signaling factor levels had no adverse effects on patients' liver or kidney function,or blood routine examinations.CONCLUSION:Perilla oil treatment is safe in clinical use,can regulate blood lipid levels and protects the function of endothelial cells.

Coronary microembolization induced myocardial contractile dysfunction and tumor necrosis factor-α mRNA expression partly inhibited by SB203580 through a p38 mitogen-activated protein kinase pathway

Background The microemboli produced during spontaneous plaque rupture and ulceration and during coronary intervention will reduce coronary reserve and cause cardiac dysfunction. It is though that inflammation caused by the microinfarction induced by the microembolization may play an essential role. It is known that the activation of p38 mitogen-activated protein kinases （MAPK） in both infected and non-infected inflammation in myocardium may cause a contractile dysfunction. But the relation between the activation of p38 MAPK and microembolization is still unknown. Methods Sprague-Dawley rats were randomly divided into three groups： Sham group, coronary microembolization （CME） group and SB203580 group （n=-10 per group）. CME rats were produced by injection of 42 pm microspheres into the left ventricle with occlusion of the ascending aorta. SB203580, a p38 MAPK inhibitor, was injected into the femoral vein after the injection of microspheres to make the SB203580 group. Left ventricular ejection fraction （LVEF） was determined by echocardiography. The protein concentration of P38 MAPK in the myocardium was assessed by Western blotting. The relative expression of mRNA for tumor necrosis factor （TNF）-a was assessed by the technique of semi-quantitative polymerase chain reaction amplification. Results LVEF was depressed at three hours up to 12 hours in the CME group. Increased p38 MAPK activity and TNF-a mRNA expression were observed in the CME group. The administration of SB203580 partly inhibited p38 MAPK activity, but did not fully depress the TNF-α expression, and partly preserved cardiac contractile function. Conclusions p38 MAPK is significantly activated by CME and the inhibition of p38 MAPK can partly depress the TNF-a expression and preserve cardiac contractile function.

Mitogen-activated protein kinase-activated protein kinase 2 regulates tumor necrosis factor-induced interleukin-6 expression via human antigen R

Background Human antigen R （HuR） is a ubiquitously expressed member of the ELAV family, and has relatively high cytoplasmic abundance in lung tissue regenerating after injury. In this study, we investigated whether mitogen-activated protein kinase （MAPK）-activated protein kinase 2 （MK2） and HuR participate in the tumor necrosis factor （TNF）-induced expression of interleukin-6 （IL-6）. Methods Human pulmonary microvascular endothelial cells were treated with TNF following short interfering RNAmediated knockdown of MK2 or HuR. Cell supernatants were collected to detect the mRNA and protein expression of IL-6 at different time points, The expression and half-life of IL-6 mRNA were then determined in cells that had been treated with actinomycin D. Finally, after knockdown of MK2, the cytoplasmic expression of HuR protein was analyzed using Western blotting. Results MK2 or HuR knockdown decreased both the mRNA and protein expression of IL-6 in TNF-stimulated cells. In MK2 knockdown cells, the half-life of IL-6 mRNA was reduced to 36 minutes, compared with 67 minutes in the control group. In HuR knockdown cells, the half-life of IL-6 mRNA decreased from 62 minutes to 24 minutes. Further analysis revealed that knockdown of MK2 resulted in reduced HuR protein expression in the cytoplasm. Conclusions MK2 regulates the TNF-induced expression of IL-6 by influencing the cytoplasmic levels of HuR.

关黄柏多糖对大鼠痛风性肾病的改善作用及机制研究

目的研究关黄柏多糖(PAP)对大鼠痛风性肾病(GN)的改善作用,并通过丝裂原激活蛋白激酶p38(p38 MAPK)/核因子κB(NF-κB)/肿瘤坏死因子α(TNF-α)信号通路初步探讨其作用机制。方法将60只大鼠按体重分层后随机分为正常组(水)、模型组(水)、别嘌醇组(阳性对照,20 mg/kg)和PAP高、中、低剂量组(100、50、25 mg/kg,以生药量计),每组10只。除正常组外,其余各组大鼠均采取1500 mg/kg氧嗪酸钾和100 mg/kg腺嘌呤联合灌胃14 d构建GN模型。造模成功后,各组大鼠灌胃相应药物/水,每天1次,连续28 d。末次给药后,检测大鼠肾功能相关生化指标[尿酸、肌酐(Cr)、血尿素氮(BUN)、黄嘌呤氧化酶(XOD)],观察大鼠肾组织病理形态学变化,检测大鼠肾组织中单核细胞趋化蛋白1(MCP-1)、TNF-α、白细胞介素6(IL-6)蛋白表达水平和p38MAPK、NF-κB p65蛋白的磷酸化水平。结果与正常组比较,模型组大鼠肾小管扩张,肾小球结构损坏,并伴有炎症浸润与纤维化;尿酸、Cr、BUN、XOD含量以及MCP-1、TNF-α、IL-6蛋白表达水平和p38 MAPK、NF-κB p65蛋白的磷酸化水平均显著升高(P<0.05或P<0.01)。与模型组比较,PAP各剂量组大鼠的肾组织病理症状均有不同程度改善;PAP高、中剂量组大鼠尿酸、Cr、BUN、XOD含量以及MCP-1、TNF-α、IL-6蛋白表达水平和p38 MAPK、NF-κB p65蛋白的磷酸化水平均显著降低(P<0.05或P<0.01)。结论PAP具有抗GN的作用,其机制可能与抑制p38 MAPK/NF-κB/TNF-α信号通路的激活有关。

三七总皂苷通过p38 MAPK通路抑制内毒素诱导的小胶质细胞活化

目的:探讨三七总皂苷(PNS)通过p38丝裂原活化蛋白激酶(p38 MAPK)通路对脂多糖(LPS)诱导活化的BV2小胶质细胞中肿瘤坏死因子-α(TNF-α)表达的影响。方法:将BV2小胶质细胞分为空白对照组(Control)、LPS激活组和LPS+三七总皂苷干预组(LPS+PNS)。CCK-8法检测BV2小胶质细胞的活力,确定最适合的药物干预浓度。利用Western Blot和免疫荧光检测BV2小胶质细胞中p38 MAPK和TNF-α的表达及p38 MAPK磷酸化水平(p-p38 MAPK)变化。结果:与空白对照组相比,PNS对BV2小胶质细胞的细胞活力无显著差异,最终选定100 mg/L作为药物干预浓度。Western Blot、免疫荧光结果提示,LPS激活后,BV2小胶质细胞中TNF-α的表达显著升高,p38 MAPK磷酸化水平增加(P<0.05);PNS干预后,与LPS激活组相比,TNF-α表达显著下降,p38 MAPK磷酸化水平降低(P<0.05)。使用p38 MAPK通路抑制剂(SB203580)作用后,PNS联合SB203580组(LPS+PNS+SB203580)中,TNF-α表达及p38 MAPK磷酸化水平变化与LPS+PNS组相比无显著差异(P>0.05)。此外,p38 MAPK在各组的变化无显著性差异(P>0.05)。结论:PNS可能通过p38 MAPK通路抑制活化的BV2小胶质细胞分泌的炎性因子TNF-α的表达。

莫西沙星联合比阿培南治疗老年重症肺炎患者疗效及对炎性因子肿瘤坏死因子相关激活蛋白和血管细胞黏附分子-1的影响

目的探讨莫西沙星联合比阿培南治疗老年重症肺炎患者疗效及对炎性因子、血清中肿瘤坏死因子相关激活蛋白(CD40L)和血管细胞黏附分子-1(VACM-1)的影响。方法采用前瞻性分析,选取浙江省人民医院收治的150例老年重症肺炎患者,根据随机数字表法分为2组,各75例。其中采用比阿培南治疗者为对照组,采用莫西沙星联比阿培南治疗者为联合组。比较2组患者疗效、病原菌检出率、炎性因子、血清CD40L和VACM-1水平。结果联合组临床总有效率明显高于对照组(97%与79%),治疗后致病菌检出率低于对照组(χ^(2)=21.714、4.812,P<0.05);联合组肺部体征、咳嗽、气喘消退时间较对照组明显缩短(t=4.751,P=0.018;t=4.369,P=0.023;t=5.134,P=0.007);联合组治疗后血清白细胞介素6(IL-6)、C-反应蛋白(CRP)、降钙素原(PCT)、CD40L和VACM-1水平均低于对照组(t=3.721,P=0.019;t=4.021,P=0.016;t=4.739,P=0.012;t=3.792,P=0.020;t=5.134,P=0.001);联合组与对照组不良反应发生率比较差异无统计学意义(χ^(2)=0.714,P=0.381)。结论莫西沙星联合比阿培南治疗老年重症肺炎患者疗效显著,患者炎症反应明显减轻,血清CD40L、VACM-1表达明显下调,且安全性较高,值得推荐。

子痫前期患者血清MAPK1、TNF-α及vWF水平及其与病情严重程度的关系

目的分析子痫前期患者血清丝裂原活化蛋白激酶1(MAPK1)、肿瘤坏死因子α(TNF-α)及血管性血友病因子(vWF)水平及其与病情严重程度的关系。方法纳入100例子痫前期患者及70例健康孕妇(对照组),根据病情严重程度将子痫前期患者分为重度组46例及轻度组54例。比较3组孕妇的常规指标(血清肌酐及尿素水平、血压、24 h尿蛋白含量)及血清TNF-α、MAPK1、vWF水平。结果重度组和轻度组的血清肌酐和尿素水平、收缩压、舒张压、24 h尿蛋白含量,以及血清TNF-α、MAPK1、vWF水平高于对照组,且重度组的上述指标高于轻度组(P<0.05)。结论子痫前期患者血清MAPK1、TNF-α及vWF水平升高,且与患者病情严重程度密切相关。检测以上指标或许有助于提高子痫前期的早期诊断率。

膝胸位中药保留灌肠治疗湿热下注型溃疡性结肠炎的临床研究

目的探讨膝胸位中药保留灌肠治疗湿热下注型溃疡性结肠炎的应用价值。方法选取2021年1月至2022年11月江西中医药大学附属医院接诊的80例湿热下注型溃疡性结肠炎患者作为研究对象,采用随机数字表法将其分为对照组和观察组,每组各40例。对照组采用侧卧位保留灌肠,观察组采用膝胸位保留灌肠,持续治疗8周。比较两组患者的临床疗效、中医证候评分、炎症因子[C反应蛋白(CRP)肿瘤坏死因子-α(TNF-α)]及凝血指标[D-二聚体(D-D)活化部分凝血酶原时间(APTT)、纤维蛋白原(FIB)],比较两组患者治疗期间不良反应发生情况。结果观察组患者的总有效率高于对照组,差异有统计学意义(P<0.05)。观察组患者治疗后的中医证候评分低于对照组,差异有统计学意义(P<0.05)。观察组患者治疗后的CRP、TNF-α、D-D、FIB均低于对照组,APTT高于对照组,差异有统计学意义(P<0.05)。两组患者不良反应总发生率比较,差异无统计学意义(P>0.05)。结论膝胸位中药保留灌肠治疗湿热下注型溃疡性结肠炎患者效果显著,可有效减轻患者临床症状,下调炎症指标,改善凝血功能。

金合欢素对丝裂原蛋白激酶p38α活性抑制作用的研究

目的探讨金合欢素对丝裂原蛋白激酶p38α(p38αMAPK)的抑制作用。方法应用分子对接软件Autodock Vina将金合欢素与靶酶三维晶体结构1KV2的活性位点进行分子对接,而后利用脂多糖诱导的小鼠RAW 264.7细胞炎症模型验证金合欢素对p38αMAPK的抑制作用。结果金合欢素可结合于1KV2的活性位点处而阻碍底物三磷腺苷的进入,对p38αMAPK的生物学功能产生抑制作用。与模型组相比,阳性对照药地塞米松和金合欢素不同浓度干预后可明显下调小鼠RAW 264.7细胞炎症模型细胞上清液中一氧化氮和肿瘤坏死因子-α水平(P<0.05)。结论金合欢素可能是通过作用于p38αMAPK活性位点而发挥抗炎的药理活性。

重组人源肿瘤坏死因子受体融合蛋白对大鼠下颌骨缺损的干预效果

目的:通过大鼠下颌骨缺损再植的实验动物模型,基于Janus激酶-信号转导与转录激活因子(Janus kinase-signal transducers and activators of transcription,JAK-STAT)通路探讨注射重组人源肿瘤坏死因子受体融合蛋白(recombinant human tumor necrosis factor receptor:Fc fusion protein,rhTNFR:Fc)对下颌骨缺损大鼠的干预效果。方法:将70只(雌雄各半)7~8周龄的健康SD大鼠随机分为3组,分别为空白组(10只)、对照组(30只)和实验组(30只)。实验组:于大鼠骨缺损处注入含有Rh TNFR:Fc(45 mg/kg)的Bio-Oss骨颗粒,其上覆盖Bio-Gide胶原膜;对照组:于大鼠缺损处注入含25μL 0.9%氯化钠溶液的Bio-Oss骨颗粒,同样覆盖Bio-Gide胶原膜;空白对照组为健康大鼠。取所有大鼠下颌骨组织,观察其形态学变化,行组织形态计量学测定。实验组和对照组大鼠于术后3、6、9周时被分批处死,取其左侧下颌骨进行炎症因子白细胞介素-6(interleukin-6,IL-6)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)表达水平的检测,同时检测组织中凋亡相关分子Bax、Bcl-2、p-JAK2和p-STAT3蛋白的表达。结果:与对照组比较,实验组的缺损面积显著减少,IL-6和TNF-α的浓度显著下降,而凋亡相关分子Bax、p-JAK2和p-STAT3的表达显著下降,Bcl-2表达显著升高(P<0.05);与空白组比较,对照组和实验组缺损面积显著增加,缺损模型建立成功。结论:局部注射rh TNFR:Fc与Bio-Oss骨颗粒能够有效地促进下颌骨缺损修复,并且效果显著优于只使用Bio-Oss骨颗粒的组别,其修复作用与抑制细胞凋亡的能力相关,通过实验我们发现JAK2-STAT3细胞信号通路在其中发挥了重要作用。

大柴胡汤联合生长抑素治疗急性胰腺炎的效果及对血清炎性指标、生化指标的影响

目的 探讨大柴胡汤联合生长抑素治疗急性胰腺炎(AP)患者效果及对血清炎性指标、生化指标等的影响。方法 选取2019年1月—2022年12月收治的AP 148例,按照治疗方法不同将其分为观察组和对照组2组各74例,观察组采用大柴胡汤联合生长抑素治疗,对照组采用生长抑素治疗。比较2组治疗后临床效果,治疗前后中医症状积分、血清炎性指标[降钙素原(PCT)、肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)]、生化指标(血钙、血淀粉酶、尿淀粉酶)及外周血单核细胞p38丝裂原活化蛋白激酶(p38MAPK)、磷酸化-丝裂原活化蛋白激酶p38抗体(p-p38MAPK),以及治疗期间不良反应。结果 治疗后,观察组总有效率91.89%(68/74)高于对照组77.03%(57/74)(P<0.05)。治疗后,2组各中医症状积分、血清炎性指标、血淀粉酶、尿淀粉酶及外周血单核细胞p38MAPK、p-p38MAPK均低于治疗前;观察组血钙高于治疗前和对照组,各中医症状积分、血清炎性指标、血淀粉酶、尿淀粉酶及外周血单核细胞p38MAPK、p-p38MAPK低于对照组(P<0.01)。治疗期间,2组不良反应总发生率比较差异无统计学意义(P>0.05)。结论 大柴胡汤联合生长抑素治疗AP患者效果显著,可减轻炎症反应,调节消化酶水平,下调外周血单核细胞p38MAPK、p-p38MAPK表达水平,进而改善临床症状。

别孕烯醇酮对老年雄性小鼠术后谵妄的防治作用及其机制

目的 观察别孕烯醇酮(Allopregnanolone,Allo)腹腔皮下注射对老年雄性小鼠术后谵妄(POD)的防治作用,并探讨其可能作用机制。方法 96只6月龄雄性C57BL/6J小鼠随机分为A、B、C组及对照组(各24只):对照组小鼠正常饲养,A、B、C组小鼠行右下肢胫骨骨折内固定术(常用POD动物模型制备方法),A组小鼠术后1小时腹腔注射别孕烯醇酮8 mg/kg,术后第6、24、48小时分别皮下注射8 mg/kg的别孕烯醇酮,B组小鼠术后1小时腹腔注射等体积溶剂2-羟丙基-β-环糊精,术后第6、24、48小时分别皮下注射等体积溶剂2-羟丙基-β-环糊精,C组小鼠术后未予任何药物。分别于术前及术后第1、3、7天采用Morris水迷宫实验评估小鼠空间学习和记忆能力,测试结束后各组随机处死6只小鼠,取海马组织检测各组小鼠海马组织炎性因子及磷酸化p38丝裂原活化蛋白激酶(p-p38MAPK)、核因子κB(NF-κB)、NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor protein 3,NLRP3)通路相关蛋白。结果 与对照组比较,术后第1、3、7天C组小鼠逃避潜伏期长、平台穿越次数少(P均<0.05);与C组比较,术后第1、3、7天A组小鼠逃避潜伏期短、平台穿越次数多(P均<0.05)。与对照组比较,术后第1、3、7天C组小鼠海马组织肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)、p-p38MAPK、NF-κB、NLRP3相对表达量高(P均<0.05);与C组比较,术后第1、3、7天A组小鼠海马组织TNF-α、IL-1β、p-p38MAPK、NF-κB、NLRP3相对表达量低(P均<0.05)。结论 别孕烯醇酮腹腔及皮下注射可减轻老年雄性小鼠右下肢胫骨骨折内固定术后POD,海马组织TNF-α、IL-1β、p-p38MAPK、NF-κB、NLRP3表达低,其机制可能为别孕烯醇酮可抑制海马组织TNF-α、IL-1β、p-p38MAPK、NF-κB、NLRP3表达。

血清IL-2、IL-6、IL-8、TNF-α、CRP在儿童IBD早期诊断及活动性评估中的应用研究

目的:分析血清白介素-2(IL-2)、白介素-6(IL-6)、白介素-8(IL-8)、肿瘤坏死因子-α(TNF-α)、C反应蛋白(CRP)在儿童炎症性肠病(IBD)中的早期诊断效果及活动性评估作用。方法:将2020年2月—2023年2月江西省儿童医院收治的108例IBD患儿[溃疡性结肠炎(UC)、克罗恩病(CD)]纳入观察组,另将同期于本院体检的84例健康儿童纳入对照组。采集两组静脉血,分析对比观察组与对照组IL-2、IL-6、IL-8、TNF-α、CRP的差异;依据小儿溃疡性结肠炎活动指数(PUCAI)、小儿克罗恩病活动指数(PCDAI)将UC、CD患儿分为不活动组与活动组,对比不活动组、活动组与对照组的IL-2、IL-6、IL-8、TNF-α、CRP差异;另绘制受试者操作特征(ROC)曲线,分析IL-2、IL-6、IL-8、TNF-α、CRP检测诊断IBD的效能。结果:观察组的IL-2、IL-6、IL-8、TNF-α、CRP均高于对照组,差异均有统计学意义(P<0.05);UC活动组的IL-2、IL-6、IL-8、TNF-α、CRP均高于UC不活动组与对照组,CD活动组的IL-2、IL-6、IL-8、TNF-α、CRP均高于CD不活动组与对照组,差异均有统计学意义(P<0.05);ROC结果显示:IL-2、IL-6、IL-8、TNF-α、CRP联合检测诊断IBD的曲线下面积(AUC)高于单独检测。结论:IL-2、IL-6、IL-8、TNF-α、CRP在IBD患儿内呈异常表达,且处在活动期的患儿,其水平均更高,上述指标联合检测可早期及时诊断出IBD。

益肾活血汤辅助治疗对维持性血液透析患者炎性因子、高凝状态及血管内皮功能的影响

目的 探讨益肾活血汤辅助治疗对维持性血液透析患者炎性因子、高凝状态及血管内皮功能的影响。方法 选取2019年6月—2022年12月收治的慢性肾衰竭尿毒症200例,根据治疗方式不同将其分为观察组(采用益肾活血汤辅助维持性血液透析治疗,102例)和对照组(采用维持性血液透析治疗,98例)2组。比较2组治疗后临床效果,治疗前后炎性因子、凝血功能和血管内皮功能,以及治疗期间不良反应。结果 治疗后,观察组总有效率(93/102,91.18%)高于对照组(68/98,69.39%)(P<0.01)。治疗后,2组C反应蛋白、肿瘤坏死因子-α、白细胞介素-6、凝血酶原时间、活化部分凝血活酶时间及内皮素-1均较治疗前降低,且观察组低于对照组;硫化血红蛋白、一氧化氮均较治疗前升高,且观察组高于对照组(P<0.01)。治疗期间,2组不良反应发生率比较差异无统计学意义(P>0.05)。结论 采用益肾活血汤辅助治疗维持性血液透析患者能改善炎性因子水平、机体高凝状态及血管内皮功能,且安全性较高。

Characterization of a small molecule inhibitor of tumor necrosis factor-alpha production

Background Numerous studies have shown that reducing the level of tumor necrosis factor-alpha （TNFα） through the use of anti-TNF antibodies or soluble TNF receptor is a safe and efficacious treatment to inflammatory diseases such as rheumatoid arthritis. Therefore, novel approaches to achieve this outcome are desired. The aim of this study was to investigate the characterization of a small molecule inhibitor, Y316, which blocks TNF mRNA upregulation and TNF production by lipopolysaccharides （LPS） stimulated monocytes. Methods Peripheral blood mononuclear cells （PBMC） from healthy volunteers were plated in 24-well plates and stimulated with LPS （1 pg/ml）, phorbol-12-myristate-13-acetate （PMA） （100 ng/ml）, zymosan （10 IJg/ml） and Tsst （100 ng/ml）. Supernatants were collected after 4-hour culture at 37℃, and quantitative determination of TNFα, interleukin-113 （IL-1β）, IL-6, IL-8, IL-10 and IL-2 production in the supernatants was performed by colorimetric enzyme-linked immunosorbent assay （ELISA）. Total RNA of PBMC was isolated and cytokine mRNA quantitation was performed by using a RNA level measuring kit （R ＆ D Systems）. PBMC were pretreated with Y316 （10 μmol/L, 1 μmol/L, 0.1 μmol/L, 0.01 μmol/L and 0.001 μmOl/L） or dimethyl sulfoxide at 37℃ for 10 minutes, and then stimulated with LPS or PMA, protein concentrations of p44.42, IKBa, P38 and Jun NH2-terminat kinase were determined by Western blotting. Cyclic adenosine-3＇,5＇-monophosphate （cAMP） of PBMC was measured by enzyme immunoassay kit （Amersham Pharmacia Biotech）. Results Y316 blocked TNF production and inhibited the upregulation of TNF mRNA levels in response to LPS, and also prevented the production of IL-1 and IL-6. In contrast, Y316 augmented the production of IL-10 in LPS-stimulated monocytes. Y316 failed to prevent the production of IL-2 and TNF in antigen-stimulated T cells, suggesting that its effects may be cell-type specific. Y316 prevented the phosphorylation and activation of the MAPK, ERK, and therefore appeared to mediate its effects on TNF by acting at an early point in the signaling cascade induced in response to LPS. There was no effect of Y316 on cAMP levels either alone or in the presence of LPS. Conclusions Y316 appears to be a small molecule inhibiting TNF production, which may act via a novel mechanism. Identification of the target of Y316 may lead to the development of alternative strategies for achieving selective cytokine inhibition.

p38丝裂原活化蛋白激酶抑制剂对脓毒症大鼠多器官损伤的保护作用研究

目的 探讨脓毒症致多器官损伤的原因,以及p38丝裂原活化蛋白激酶(MAPK)抑制剂的保护作用和机制。方法 采用盲肠结扎穿刺术(CL P)制备脓毒症大鼠模型,治疗组采用术前给予p38MAPK抑制剂SB2 0 35 80灌胃。在不同时间点观察大鼠血清肿瘤坏死因子- α(TNF-α)和白细胞介素- 1β(IL- 1β)以及生化指标如丙氨酸转氨酶(AL T)、尿素氮(BUN)、肌酐(Cr)、肌酸磷酸激酶同工酶(CPK -MB)浓度的变化。结果 CL P术后大鼠血清TNF-α、IL -1β显著升高,AL T、BU N、Cr、CPK- MB也进行性升高;血清AL T、BU N、Cr、CPK -MB变化与TNFα、IL 1β呈显著正相关。应用SB2 0 35 80后,血清TNF-α、IL- 1浓度显著降低,同时AL T、BUN、Cr、CPK MB也降低。结论 TNF-α、IL- 1β的大量释放是脓毒症致多器官损伤的原因之一,通过调控p38MAPK信号转导通路可对脓毒症所致多器官损伤起保护作用。

STAT3与MAPK蛋白协同调节肿瘤坏死因子-α转录活性

探讨信号转导和转录激活因子3(signal transducer and activator of transcription 3,STAT3)与丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)在体内是否存在相互作用,并观察其作用如何影响肿瘤坏死因子-α(tumor necrosis factor,TNF-α)的转录活性.采用聚合酶链反应技术,从人Flag-p38和Flag-细胞外信号蛋白调节激酶2(extracellular-signal regulated protein kinase 2,ERK2)中扩增出p38和ERK2基因,将其插入载体pcDNA3-HA中;用Westernblot方法在293T细胞中检测其表达后应用免疫共沉淀技术检测STAT3蛋白与p38/ERK2蛋白之间是否存在相互作用.然后应用报告基因技术检测这种相互作用如何影响TNF-α的转录表达,并在应用RNA干扰技术将STAT3通路抑制后,观察TNF-α启动子转录活性如何变化.酶切和测序结果表明,扩增的p38和ERK2基因序列正确,大小为1080bp,在293T细胞中正确表达大小约40ku的p38和ERK2蛋白.免疫共沉淀实验结果证实,p38和STAT3蛋白以及ERK2和STAT3蛋白在体内相互作用.下游基因TNF-α荧光素酶活性实验显示,p38和STAT3蛋白以及ERK2和STAT3蛋白复合物协同升高TNF-α的活性,应用STAT3的干扰RNA后其活性则明显下降.该研究表明,STAT3和p38/ERK2蛋白在体内存在相互作用,STAT3与p38、STAT3与ERK2均对TNF-α的表达发挥协同效应,在阻断STAT3通路后,STAT3与p38、STAT3与ERK2对TNF-α表达的协同效应将明显降低.